lncRNA FGD5-AS1靶向miR-15a-5p调控LPS诱导的脓毒症细胞损伤

lncRNA FGD5-AS1 targets miR-15a-5p to regulate LPS-induced sepsis cell injury

目的:探讨长链非编码RNA FGD5-AS1(lncRNA FGD5-AS1)对LPS诱导的脓毒症细胞损伤的影响及分子机制。方法:500 ng/ml脂多糖(LPS)诱导THP-1细胞损伤。THP-1细胞分为对照组、LPS组、pcDNA组、pcDNA-FGD5-AS1组、anti-miR-NC组、anti-miR-15a-5p组、pcDNA-FGD5-AS1+miR-NC组、pcDNA-FGD5-AS1+miR-15a-5p组。RT-qPCR检测lncRNA FGD5-AS1和miR-15a-5p表达;流式细胞术检测细胞凋亡;Western blot法检测蛋白表达;ELISA检测IL-6、IL-1β、TNF-α水平。双荧光素酶报告实验检测lncRNA FGD5-AS1和miR-15a-5p的靶向关系。结果:与对照组相比,经LPS诱导的THP-1细胞中,lncRNA FGD5-AS1表达降低,miR-15a-5p表达升高(P<0.05)。过表达lncRNA FGD5-AS1和下调miR-15a-5p均能够显著降低细胞凋亡以及炎症细胞因子IL-6、IL-1β、TNF-α表达(P<0.05)。lncRNA FGD5-AS1靶向调控miR-15a-5p表达,上调miR-15a-5p能逆转过表达lncRNA FGD5-AS1对LPS诱导的THP-1细胞凋亡以及炎症细胞因子IL-6、IL-1β、TNF-α的影响(P<0.05)。结论:过表达lncRNA FGD5-AS1通过靶向下调miR-15a-5p表达减轻LPS诱导的THP-1细胞损伤。

Objective:To investigate effect and molecular mechanism of long non-coding RNA FGD5-AS1(lncRNA FGD5-AS1)on LPS-induced sepsis cell injury.Methods:Injury of THP-1 cells was induced by 500 ng/ml LPS.THP-1 cell were divided into control group,LPS group,pcDNA group,pcDNA-FGD5-AS1 group,anti-miR-NC group,anti-miR-15a-5p group,pcDNA-FGD5-AS1+miR-NC group and pcDNA-FGD5-AS1+miR-15a-5p group.RT-qPCR was used to detect expressions of lncRNA FGD5-AS1 and miR-15a-5p;flow cytometry was used to detect cell apoptosis;Western blot was used to detect protein expressions;ELISA was used to evaluate levels of IL-6,IL-1β,TNF-α.Targeted relationship between lncRNA FGD5-AS1 and miR-15a-5p was detected by double luciferase reporting experiment.Results:Compared with control group,expression of lncRNA FGD5-AS1 was decreased and miR-15a-5p expression was increased in LPS-induced THP-1 cells(P<0.05).Overexpression of lncRNA FGD5-AS1 and down-regulation of miR-15a-5p can significantly reduce apoptosis and expressions of inflammatory cytokines IL-6,IL-1βand TNF-α(P<0.05).lncRNA FGD5-AS1 targeted to regulate expression of miR-15a-5p,up-regulating of miR-15a-5p can reverse effect of lncRNA FGD5-AS1 overexpression on LPS-induced apoptosis and expressions of inflammatory cytokines IL-6,IL-1βand TNF-α(P<0.05)of THP-1 cells.Conclusion:Overexpression of lncRNA FGD5-AS1 attenuates LPS-induced THP-1 cell injury by targeting down-regulating miR-15a-5p.